CN107459416B

CN107459416B - Organic fertilizer and components thereof

Info

Publication number: CN107459416B
Application number: CN201710332054.1A
Authority: CN
Inventors: G·E·哈曼; X·雷
Original assignee: Cornell University
Current assignee: Cornell University
Priority date: 2011-11-23
Filing date: 2012-11-21
Publication date: 2021-10-22
Anticipated expiration: 2032-11-21
Also published as: CN104203871A; US20140323297A1; WO2013078365A1; US9249061B2; CN107459416A

Abstract

The present invention relates to a fertilizer comprising at least one microorganism selected from the group consisting of trichoderma viride (NRRL B-50520), scoparia glauca (NRRL B-50521), scoparia glauca (NRRL B-50522), and combinations thereof; and a substrate that is acted upon by the at least one microorganism to release nitrogen. The invention also relates to a fertilizer comprising at least one microorganism selected from keratin degrading microorganisms and a keratin substrate acted on by the at least one microorganism to release nitrogen. The invention also relates to a method of using the fertilizer to enhance plant growth, and to a method of manufacturing a fertilizer. The invention also relates to an isolated strain of Trichoderma viride (NRRL B-50520) and two isolated strains of Gliocladium brevicaulis (NRRL B-50521, NRRL B-50522).

Description

Organic fertilizer and components thereof

The application is a divisional application of Chinese application with application number 201280062108.2, application date 2012 and 11/21, entitled "efficient organic fertilizer and components thereof", and the priority of U.S. provisional application No. 61/563,355 filed 2011 and 11/23, the entire contents of which are incorporated herein by reference.

The subject matter of the present application was made with the support of approval number 0945724 from the national science foundation of the united states. The government has certain rights in this invention.

Technical Field

The invention relates to a high-efficiency organic fertilizer and components thereof.

Background

Fertilizers used in lawn and other applications often consist of inorganic mixtures of salts that provide nitrogen, phosphorus and potassium. Fertilizers for different uses may be liquid or solid and contain a wide range of nitrogen to phosphorus to potassium ("N: P: K"), as well as other plant nutrients depending on the intended use of the product. In general, fertilizer products are composed mainly of salts of inorganic compounds, and are therefore unsuitable for the organic state.

Many conventional fertilizer products are very well functioning. However, these fertilizer products are of increasing interest because they release high levels of nitrates into groundwater resulting in contaminated water. A fertilizer product has a composition of 32: 2: 8 (% N: P)₂O：K₂O₅) The formulation of (1). Analysis of this package showed that of these 32%, 3.8% were ammonium compounds, 53% were urea, 39% were water soluble N, and 3.8% were other N. If these compounds are not absorbed by the plants, nitrate and urea are immediately available for infiltration into the groundwater. Unfortunately, plants typically Use only about 33% of the total nitrogen fertilizer applied (Amall et al, "Relationship shift Between the greenhouse nitrogen Use Efficiency and Response Index in Winter Wheat," J.plant Nutr.32: 502-515(2009)), while the remainder can be metabolized to nitrogen oxides, which are potent greenhouse gases, or which are impregnated into soil and surface water in the form of nitrates and nitrites where they can be toxic (EPA limits for drinking water are 10ppm and 1ppm, respectively), and where they can produce decay of algae and other microorganisms to create anaerobic areas where plants, fish and other dwelling animals cannot survive. There is a need for fertilizers that do not have these adverse environmental effects.

Due to regulatory concerns about eutrophication of water bodies, fertilizers for lawn use are being improved. For this reason, phosphorus-containing lawn fertilizers are banned in some places. Nitrate is a contaminant at least as severe as phosphorus, and many places, such as the long island, are intended to limit water contamination from this source. The severity of the problem can be explained from the following citations on Chesapekey gulf (Dewar et al, "Urban Fertilizers and the Chesapeak Bay: An Opportunity for Major polarization Reduction," Executive Summary, environmental market, Research and Policy Center (2011)):

over 26 years, attempts have been made in various states of the chesapercha bay to clean the bay, but it remains clogged with extremely harmful excess contamination. To achieve a clean, sustainable bay, the nitrogen levels in bay waters must be reduced by an additional 30% and phosphorus reduced by an additional 8% in each state of the bay basin-although a 30% population growth is expected by 2030. Such a reduction in size is only possible if the government targets all sources of nutrient contamination of the watershed.

Excess nitrogen and phosphorus, as well as sediments, are a major cause of often poor water quality in the gulf and the waters into which they flow. Approximately 30% of the phosphorus load in the gulf comes from urban and suburban runoff. Those same developed lands account for 10% of nitrogen contaminated runoff. However, measures to reduce nutrient runoff from developed land are far from adequate.

While the state regulatory body does a better job requiring farmers to control nutrient-laden runoff from the respective fields, the state largely neglects the major crops of the watershed: and (4) grass. Throughout the gulf basin, approximately 380 ten thousand acres are now growing turfgrass, and as residential developments expand and replace farmlands, the growing area is also growing. Turfgrass is by far the largest crop in maryland, growing as much as 130 ten thousand acres throughout the state. In contrast, in 2009, the area of all other crops planted in maryland was 150 ten thousand acres. However, it is the least regulated crop of the major crops in the state.

Turfgrass becomes a pollution problem when it is covered with too much fertilizer containing nitrogen and phosphorus. The nutrients in the fertiliser may help maintain a healthy lawn, but when excessive, they may wash into nearby bodies of water when it rains or snows. Excess fertilizer nutrients may also directly penetrate into groundwater. Whether the fertilizer is organic or chemical, its nutrients can damage the gulf and local waterways.

Tracking fertilizer usage on developed land is so much of no concern that the state is not counting it, but records from the Maryland department of agriculture show that "non-agricultural use" fertilizer sales are rapidly overtaking farm fertilizer sales. The best estimates indicate that land owners in maryland apply at least 8600 thousand pounds of nitrogen fertilizer per year on lawns in the state.

This fertilizer enters rivers and bays. In a watershed in the suburb of bald, researchers found that 56% of the nutrients in a local river came from lawn fertilizer. Scientists in texas, wisconsin, minnesota, connecticut and canada have also demonstrated that contaminants in lawn fertilizer can significantly compromise surface water quality.

As in 1-2 above.

Several regulations in the long island require that the committee urge all citizens to voluntarily avoid the use of chemical fertilizers and lawn pesticides and urge the use of organic lawn care "because the health of children and citizens in [ towns ], their water, environment and the long island strait would benefit from reducing the use of chemical fertilizers and lawn pesticides. Typical municipal regulations and regulations for pesticides/herbicides, connecticut state river consortium. There is a need and potential market for organic turf fertilizers with a lower potential for water contamination. For example, the above regulations do not require the use of only organic fertilizers, primarily because there are no pollution-free, organic and cost-effective products with sufficient performance.

The synthetic fertilizers used primarily are very effective in providing quality turf, but they are environmentally damaging. Unfortunately, as noted above, organic fertilizers are not themselves prodigiosin. They may also contaminate water areas because they still require equal amounts of nitrogen to provide a vigorous, luxurious lawn. Generally, current organic fertilizers have serious drawbacks. These include the inability to provide sufficiently thriving turf shortly after application, they typically contain only 7 to 9% of any form of N. This means that they do not provide a sufficient amount of N for luxurious green lawns unless high ratios are used, since the efficacy of use depends on the dosage used. In addition, in order to obtain sufficient performance,they must be present in a 2-6 fold ratio

Application rate. Thus, a bag weighing about 37 pounds

Fertilizers were fertilized at 15,000 square feet for 1-2 months (the company advocates four applications per growing season); the equivalent result of using a low N organic fertilizer would require about 3 bags to provide the same amount of N. These factors make conventional organic fertilizers inconvenient to use, and customers generally do not wish to carry and broadcast 100-.

The fact that such a large amount needs to be applied makes the conventional organic fertilizer uneconomical. Even if a bag of 25 pounds of organic fertilizer is slightly less expensive than inorganic fertilizer, the total price of over 100 pounds will be much higher. Commercial inorganic fertilizers typically sell for a bag at about $ 55, while the same amount of N in most organic fertilizers will cost over $ 100.

The present invention is directed to overcoming these and other deficiencies in the art and providing answers to many of the problems of traditional fertilizers while providing several different paths for new and unique products that will meet the needs of users and limit environmental issues.

Summary of The Invention

The present invention relates to a fertilizer comprising at least one microorganism selected from the group consisting of: trichoderma viride (Trichoderma viride) (NRRL B-50520), Scopulariopsis brevicaulis (NRRL B-50521), Scopulariopsis brevicaulis (NRRL B-50522), and combinations thereof. The fertilizer also includes a substrate that is acted upon by at least one microorganism to release nitrogen.

Another aspect of the invention relates to a method of enhancing plant growth. The method comprises providing a fertilizer comprising at least one microorganism selected from the group consisting of: trichoderma viride (NRRL B-50520), Gliocladium brevicaulis (NRRL B-50521), Gliocladium brevicaulis (NRRL B-50522), and combinations thereof; and a substrate that is acted upon by at least one microorganism to release nitrogen. The method further comprises contacting the fertilizer with the plant or plant seed under conditions effective to increase the growth of the plant or plant seed compared to a plant or plant seed to which the fertilizer is not applied.

The invention also relates to a fertilizer comprising at least one microorganism selected from keratin degrading microorganisms and a keratin substrate acted on by the at least one microorganism to release nitrogen.

Another aspect of the invention relates to a method of making a fertilizer comprising an amino acid and ammonia. The method comprises providing at least one microorganism selected from the group consisting of: trichoderma viride (NRRL B-50520), Gliocladium brevicaulis (NRRL B-50521), Gliocladium brevicaulis (NRRL B-50522), and combinations thereof and provides a substrate that is acted upon by at least one microorganism to release nitrogen. The method further comprises contacting the at least one microorganism with the substrate under conditions effective to cause the at least one microorganism to exert an action on the substrate and produce a liquid suspension comprising the amino acid and ammonia.

The invention also relates to an isolated Trichoderma viride strain deposited at the Agricultural Research Service Culture Collection (Agricultural Research Service Culture Collection) under accession number NRRL B-50520.

Another aspect of the invention relates to an isolated strain of Scopulariopsis brevicaulis deposited at the agricultural research service culture Collection under the accession number NRRL B-50521.

Another aspect of the invention relates to an isolated strain of Scopulariopsis brevicaulis deposited at the agricultural research service culture Collection under the accession number NRRL B-50522.

The present invention relates to the development of a highly efficient organic fertilizer that can be used for a variety of purposes and that can minimize or eliminate groundwater contamination from nitrates. It includes the identification of slow release nitrogen forms and methods of use thereof. It also includes microbial agents that can degrade complex forms of nitrogen, particularly proteins produced from agricultural waste products, and release plant-available nitrogen into the soil or into fermenters. It describes the use of microorganisms that are endophytic plant symbionts that enhance the activity of the fertilizers described herein and also improve plant growth and performance. The fertilizer is an effective method of delivering these organisms.

There are several requirements for a fertilizer that is acceptable to the user and non-polluting. First, there is a need for a generally applicable mechanism by which plants can more efficiently assimilate N fertilizers. One way to accomplish this is through the use of endophytic symbiotic fungi or bacteria that increase the ability of plants to use nitrogen fertilizers more efficiently ("nitrogen use efficiency" or "NUE"). NUE can be induced by increasing plant root growth and by physiological changes in the plant. Many microorganisms are potentially useful in this regard. For example, these microorganisms include bacteria designated plant growth-promoting rhizobacteria ("PGPR"), basidiomycetes (basidiomycetes) pyricularia indica (Piriformospora indica), and mycorrhizal fungi, such as those in the genus Glomus sacculus (Glomus). Other examples include Trichoderma harzianum (T.harzianum) strain RR17Bc (ATCC accession No. PTA 9708), Trichoderma harzianum F11Bab (ATCC accession No. PTA 9709), Trichoderma atroviride (T.atroviride) strain WW10TC4(ATCC PTA accession No. 9707), and Trichoderma viride (T.virens) strain 41(ATCC accession No. 20476)). These same fungi also induce many other beneficial changes in plants such as disease resistance, tolerance to abiotic stresses such as drought and salt, and increase the ability of plants to undergo photosynthesis, thereby providing other benefits to plants.

For turf, the present invention provides a functional equivalent of the assay 30: 2: 8 (N: P)₂O₅∶K₂O) is an organic fertilizer with an analysis of 12-16: 0-10: 5-15. A prototype process to produce a granulated fertilizer suitable for dispersion in a standard drop or broadcast fertilizer spreader, which fertilizer comprises the organisms claimed in the present invention, is also described. Also described is a method for preparing a powder formulation which can be incorporated into potting media, applied to furrows and the like, and also a process for preparing a liquid organic fertilizer. The result is a new fertilizer, which can be classified as organic.

Another useful property that may be provided by a component of the fertilizer is the slow release of nutrients. Fertilizers consisting of only soluble salts rapidly release their nutrients and allow the nutrients to be flushed into the ground water, which quickly becomes a contaminant. As described herein, microorganisms, particularly selected Trichoderma (Trichoderma) strains, can significantly promote plant growth, improve abiotic stress tolerance, increase plant nitrogen use efficiency, increase root growth and development, and significantly enhance general plant performance. It would be highly advantageous to incorporate such beneficial microorganisms directly onto or into fertilizers, so that the benefits of the fertilizer and beneficial microorganisms can be conveniently applied simultaneously and in the same preparation. However, the high salt content of most synthetic fertilizers will severely damage or kill the applied microorganisms contained in or on the fertilizer. The invention describes a fertilizer and a method for producing fertilizer granules consisting essentially of proteins which are not harmful to microbial agents. Contamination is particularly likely to occur if the fertiliser is applied in excess, if there is excess rain or irrigation water, or when the fertiliser is applied to sandy and nutrient leaching soils. Since it is not foreseen where or how to apply the fertilizer, nitrogen may be in a slow release form, enabling the plant to absorb more nutrients with the slow release of the applied nutrients, rather than immediately. There are two basic processing methods. First, the standard process in the industry is to produce nitrogen in various forms that slowly release nitrogen. There are many slow-release forms of nitrogen in commercial commerce today. These forms include urea formaldehyde, methylene urea, sulfur coated urea (urea), and others. These are well known in the art, see, for example, U.S. patent 8,182,572 to King et al, which is hereby incorporated by reference in its entirety. Secondly, there are various complex forms of nitrogen, usually in the form of protein, manure or compost. These materials slowly release their own nitrogen due to microbial activity.

"organic" materials can be certified and regulations relating to them are institutional dependent. The organic materials institute ("OMRI") certifies products according to its own specifications. OMRI is a private organization with perfect authentication. Many, but not all, agricultural waste products, such as compost, manure, etc., can be certified, but their manufacturing requirements are relatively stringent. Some well-defined chemicals may be used, but their use is strictly regulated. For example, sodium nitrate is only acceptable when it is in mined form (known as chilium nitrate), rather than manufactured, and its use is limited even for such materials. Additionally, the near organic certification is the USDA bioprefred product listing. Products with this name must be derived from bio-based agricultural products at a certain percentage of the total. The minimum content of farm-derived products of fertilizer is 71%. This may include many of the same materials listed by OMRI, with exceptions. For example, peat moss is eligible for OMRI certification but not certified by the bioprefred program. Compost, manure, protein sources from farm animals and plants are acceptable for both. One significant difference is that the OMRI registered products must be 100% of their certified products, but for bioprefred, up to 29% may be products outside the bioprefred list. Thus, some synthetic slow-release synthetic nitrogen forms are available for bioperfred products, but not for OMRI-labeled products.

The present invention describes microorganisms (i.e. fungi) that efficiently degrade keratin in feathers/feather meal. These microorganisms rapidly release the amino acids and ammonia in the feather meal into the semi-solid medium. For example, inocula of fungi have many uses and can be prepared as dry spore formulations or liquid formulations. The invention also describes a method for producing granules capable of being broadcast, with or without a coating of microorganisms, or with or without microorganisms incorporated into the granules, which provide a slow-release microbial activated release of nitrogen for plant growth, especially in combination with other beneficial fungi. The incorporation of the microbially activated nitrogen system releases little or no nitrate into the soil and, therefore, little or no nitrate will be released into the groundwater. This environmental advantage is even greater if the microorganism releases N in combination with other microorganisms that can increase root growth and improve nitrogen utilization efficiency of the plant. Furthermore, the same invention, with minor modifications, can be used to produce water soluble sources or amino acids and ammonia which are highly beneficial for use as plant fertilizers.

The present invention meets the need created by traditional fertilizers by providing one or more systems comprising a fertilizer of a particular composition and function in combination with a microorganism that degrades a complex protein substrate to provide nutrients, particularly for plant growth. It also includes microorganisms (e.g., endophyte symbiota) that enhance the growth and performance of plants and provide other advantages to plant growth. The fertilizer of the present invention is an excellent delivery agent for these organisms.

The present invention is expected to provide revolutionary benefits to the fertilizer industry. First, the fertilizer of the present invention for the first time provides a fully organic or biopreferential fertilizer that performs as well as standard, widely sold inorganic fertilizers. It is also as a convenient and easy to use fertilizer as conventional inorganic fertilizers, and as a lawn fertilizer that greatly reduces nitrate contamination of groundwater and surface water. The fertilizer of the invention also provides a method for producing a new class of organic water-soluble fertilizers that contain nitrogen that is immediately available to plants in the form of amino acids and ammonia. The only product available to organic (OMRI certified) growers that contains immediately available nitrogen is chilium nitrate, and its use is limited by the OMRI regulations, which may be banned for organic use in the near future. The mixture of amino acids and ammonia will be a very useful new product, both for the ingredients of other fertilizers and as a liquid free-standing fertilizer. The present invention describes for the first time a method for producing an OMRI registrable mixture of amino acids and ammonia and the composition of such products. The focus is lawn fertiliser, which may be the most demanding material and perhaps the material used in the greatest amount, but the principles and developments are equally applicable to fertilisers for other applications. Products with these functions do not currently exist.

Brief Description of Drawings

Figure 1 shows dried granules prepared using feather powder-gelatin mixture coated with feather degrading microorganisms before and after two days of humidification. The fungus coated on the outside of the granules grew moderately. The particles are modified by the addition of a medium that supports rapid and mass growth of Trichoderma. The dried granules had a diameter of about 2 mm.

FIGS. 2A-2B show a comparison of fertilizers using formulations described in the present invention or commercially available. In FIG. 2A, the treatments used were PT (organic fertilizer, NRRL B-50521, and K1-K4, which include Trichoderma harzianum strain RR17Bc (ATCC accession PTA 9708), Trichoderma harzianum F11Bab (ATCC accession PTA 9709), Trichoderma atroviride strain WW10TC4(ATCC accession 9707), and Trichoderma viride strain 41(ATCC accession 20476)); STB

P (organic fertilizer with NRRL B-50521); and C (untreated control). In FIG. 2B, the treatments used were PT (organic fertilizer, NRRLB-50521, and K1-K4, supra); OFC (organic fertilizer granules without any fungi); and P (organic fertilizer with NRRL B-50521).

Figures 3A-3D show the harvest weight of grass over a period of time using different fertilization treatments.

Fig. 4 shows the results in sand: turf roots grown for about 6 weeks in peat mixtures and fertilized with the granules described in this example with and without beneficial microorganisms developed from a mixture of Trichoderma viride strain NRRL B-50521 and Trichoderma strain K1-K4 (Trichoderma harzianum strain RR17Bc, Trichoderma harzianum F11Bab, Trichoderma atroviride strain WW10TC4, and Trichoderma viride strain 41). The petri dish labeled 11 showed root growth on the lawn fertilized with particles without beneficial microbes. The dish labeled 12 contains a label of about 10⁵cfu/g organisms incorporated in the particles, while the 13 marked petri dishes were fertilized with beneficial fungus-coated particles in a glucan mixture (10 in 2% tapioca dextran)⁵Colony forming units (cfu)/ml to form a continuous thin coating).

Detailed Description

The present invention relates to a fertilizer comprising at least one microorganism selected from the group consisting of: trichoderma viride (NRRL B-50520), Gliocladium brevicaulis (NRRL B-50521), Gliocladium brevicaulis (NRRL B-50522) and combinations thereof. The fertilizer also includes a substrate that is acted upon by at least one microorganism to release nitrogen.

The invention may be carried out using a variety of substrates that are acted upon by microorganisms to release nitrogen, including hair, hooves, and feathers. In one embodiment, the substrate is any protein selected from the group consisting of feather, feather meal, and derivatives of urea. Any type of feather may be used, including but not limited to chicken, turkey, and duck feathers. The invention is applicable to the degradation and utilization of all substrate materials that release nitrogen.

Any number of organisms or beneficial microorganisms may be added to the fertilizer of the present invention. These organisms include those described by Harman in WO 2010/091337; harman, "Multifunctional Plant sympositions: new Tools to Enhance Plant Growth and production, "New phytol.189; 647-49 (2011); lorito et al, "Translational Research on Trichoderma: from' Omics to the Field, "annu. Rev. Phytopathol.48: 395-; shoresh et al, "Induced systematic Resistance and Plant Responses to Fungal Biocontrol Agents," Annu. Rev. phytopathohol.48: 21-43(2010), which is hereby incorporated by reference in its entirety. Other beneficial organisms include mycorrhizal fungi, plant growth-promoting rhizobacteria, azospirillum and azorhizobium. These organisms have a very large number of useful properties. Fungi alter gene expression and cause plants to exhibit increased resistance to pathogenic pathogens ("biotic stress"), exhibit increased tolerance to environmental stresses such as drought, salt, or temperature ("abiotic stress tolerance"), and increase the efficiency of nitrogen uptake ("improved nitrogen utilization efficiency [ 'NUE' ]"). The beneficial and endophytic microorganisms of the present invention can provide improved nitrogen utilization efficiency. Beneficial microorganisms can be formulated or mixed to produce a particulate or liquid suspension. These particles or liquid suspensions can be mixed directly into the soil or potting mixture. The term soil in this specification is used to include any medium capable of supporting plant growth and thus includes common soil, humus, manure, compost, sand and the like, as well as artificially produced plant growth media. The formulation is then mixed into The soil or into The volume of The planting mixture for greenhouse applications or into The upper volume of The field soil (Harman, G.E., "The Dogmas and Myths of biocontrol. changes in Perceptions Based on Research with Trichoderma harzianum T-22," Plant Dis.84: 377-393(2000), which is incorporated herein by reference in its entirety). Devices and procedures for such applications are well known and used in various agricultural industries. In one embodiment of the invention, the fertilizer further comprises a supplemental microorganism in the form of a trichoderma species. The trichoderma species may be selected from the group consisting of trichoderma virens, trichoderma harzianum, trichoderma atroviride, and combinations thereof.

Trichoderma grows among the cells in the root epidermis and cortex, and induces the surrounding plant cells to deposit cell wall material and produce phenolic compounds. This vegetative response limits Trichoderma growth in roots (see Yedidia et al, "Induction and Accumulation of PR Proteins Activity During Early Stages of Root condensation by the Mycoparasite Trichoderma harzianum Strain T-203," Plant Physiol. biochem. 38: 863. sup. 873(1999), which is incorporated herein by reference in its entirety). Endophytic plant symbiota have a much longer duration of efficacy because of their ability to grow with plants and in the environment; thus, they may have an effect for weeks or months if conditions favor them. These organisms may form on or in the roots of plants and provide benefits to the plants for at least the lifetime of an annual crop (Harman, G.E., "Myths and Dogmas of Biocontrol. changes in permissions Derived from Research on Trichoderma harzianum T-22," Plant Dis.84: 377-393(2000) and Harman et al, "Changing Paradigms on the model of Action and Uses of Trichoderma spp. for Biocontrol," outlook test Manag.19: 24-29(2008), which are incorporated herein by reference in their entirety). The formation of viable hyphae of beneficial organisms in the root cortex leads to chemical communication in the plant. In some embodiments, the fertilizer contains other supplemental microorganisms, such as those selected from the group consisting of pyricularia indica, plant rhizosphere growth-promoting bacteria, mycorrhizal fungi, and combinations thereof.

Reprogramming of plant gene expression thus occurs and brings many benefits to the plant. This ability to grow or conidium germinate on the root surface enables a variety of application methods. These application methods include, but are not limited to, seed treatment, application to soil or planting mixtures in drench that penetrate the soil volume, and in-furrow, broadcast, or spray application to the soil surface containing the roots at the time of planting. It also allows the use of very small amounts of inoculum applied in seed treatments (10s g/hectare), but subsequently results in subsequent proliferation of organisms on the root system, leading to effects that persist throughout the season, including Plant protection, greater root proliferation and enhanced root penetration into the Soil (Adams et al, "Trichoderma harzianum Rifai 1295-22media Growth Promotion of Crack (Salix fragilis) Saplings in Both clear and Metal-associated Soil," Microbial Ecol.54: 306; "2007; Harman, G.E.," Myths and Dogmas of Biocontrol. Change in microorganisms modified from Research Trichoderma Zianum T-22, "plant.84: 377; Harman et al," mutation of culture of Growth and reaction, 2008. 19. and Biocoding of plants, and 22, respectively, model of plants, 22, model of plants, 393 of fungi, and 22, avirule Plant sympositions, "Nature Rev. Microbiol.2: 43-56(2004), which are hereby incorporated by reference in their entirety).

Trichoderma strains (e.g., Trichoderma viride, Trichoderma harzianum, and Trichoderma atroviride) suitable for use in the present invention are strains that have a strong ability to colonize the root system. This ability is referred to as rhizosphere ability and is used herein to describe those organisms that are capable of colonizing the root surface or surfaces plus the surrounding volume of soil (referred to as the root surface and rhizosphere, respectively) when seeds or other point sources are applied without substantial water flow at the time of planting. Thus, the organisms of the invention have the physiological and genetic ability to proliferate in and on the root system as it develops. Rhizosphere ability is not an absolute term, and varying degrees of this ability may occur between strains (Harman, G.E., "The Development and resins of Rhizophere composite Fungi for Biological Control of Plant Pathologens," J.plant Nutrition 15: 835-843 (1992); Smith, U.S. Pat. Nos. 4,996,157 and 5,165,928, which are incorporated herein by reference in their entirety). Procedures for Measuring rhizosphere competence are known to those skilled in The art (Harman et al, "Combining Effective Strains of Trichoderma harzianum and Solid Matrix printing to Imporve Biological selected strategies," Plant diseases 73: 631. 637 (1989); Harman, G.E., "The Myths and Dogmas of Biocontrol. Change in Perceptions Based on Research with Trichoderma harzianum T-22," Plant diseases 84: 377. 393 (2000); Kloepper et al, "A Review of Issues Relationseeding catalysis of plants by Bacia," Can.J.Microol.38: 1219: 1232(1992), incorporated herein by reference in its entirety).

The fertilizer of the invention may be in the form of granules, pellets, dust, powder, slurry, film and/or liquid suspension. In one embodiment, the fertilizer is in the form of granules or pellets containing gelatin as a curing agent. The gelatin may comprise 5-20% by weight of the granules or pellets, or more preferably 7-8% by weight of the granules or pellets. In another embodiment, the fertilizer comprises at least 10^4 colony forming units per gram ("cfu") of at least one microorganism in the substrate. In yet another embodiment, the fertilizer is in the form of a liquid suspension comprising an amino acid and ammonia. The microorganisms of the present invention are growing and therefore their number is very small by weight. 10^4cfu corresponds to the proportion of microbial preparation which is only a small fraction of 10^7 in the substrate. However, the formulation can be made in many different ways, and therefore the concentration of Trichoderma or other micro-organisms in the granules or pellets can easily be varied by 100X.

The fertilizer of the present invention also contemplates supplemental sources of added nutrients. These sources include, for example, soil, water, urea, ammonium nitrate, sources that provide nitrogen, phosphorus, potassium, calcium, magnesium, sulfur and other micronutrients. At least 16 chemical elements are known to be useful in plant growth and survival. The 16 chemical elements are divided into two main classes: non-minerals and minerals. Non-mineral nutrients include hydrogen (H), oxygen (O) and carbon (C) and are present in air and water. The remaining 13 nutrients are minerals, which come from the soil, dissolve in water and are absorbed by the roots of the plants. Mineral nutrients can be further divided into two groups: macronutrients and micronutrients. Macronutrients include, but are not limited to, N, P, S, K, Ca, Mg, Na, and Si. Micronutrients include, but are not limited to, Fe, Mn, Cu, Zn, Mo, B, and Cl. Only very small (i.e. micro) amounts of micronutrients are required. These nutrients and their benefits to plants are well known to those skilled in the art. The fertilizer of the invention may also contain a soluble nitrogen source selected from the group consisting of nitrates, ammonia, ammonium salts, amino acids, urea, fish meal or extract, compost extract, seaweed extract, shrimp extract, shellfish extract, and combinations thereof.

The substrate of the invention may be blended with other sources of plant nutrients including potassium, phosphorus, iron or micronutrients.

In one embodiment, the fertilizer further comprises a non-protein binder selected from the group consisting of dextran, starch, polyvinyl chloride, and combinations thereof.

In another embodiment, the fertilizer may include a source of phosphorus selected from the group consisting of rock phosphate, sodium phosphate, potassium phosphate, bone meal, and combinations thereof.

In yet another embodiment, the fertilizer may include a potassium source selected from the group consisting of potassium chloride, potassium phosphate, potassium sulfate, new jersey green sand, organic sources of potassium, animal manure, and combinations thereof. For organic applications, sources of potassium include new jersey greensand as well as potassium sulfate and potassium chloride listed organically.

The fertilizer may alternatively comprise micronutrients selected from the group consisting of salts, substances containing iron, cobalt, manganese, magnesium, copper, calcium, boron, zinc and combinations thereof. These are readily available from a variety of sources.

Sulfur is already present in the amino acids contained in the substrate, including, for example, feather meal. Useful fertilizers can be prepared in various N: P: K ratios by mixing or adding other sources of organic substrates other than N, with or without other nutrients. In one embodiment, the fertilizer has a nitrogen content of between 10-20 wt%.

In one embodiment, the fertilizer contains a growth medium. The growth media of the present invention may include, but are not limited to, soil, sand compost, peat, rice hulls, coir (coipeat), soilless growth media containing organic and/or inorganic components, artificial plant growth substrates, polymer-based growth substrates, hydroponic nutrients and growth solutions, organic soil conditioners, water and mixtures thereof.

The fertilizer may comprise a carrier selected from the group consisting of water, aqueous solutions, slurries and powders.

Alternatively, the fertilizer may include additives such as, but not limited to, insecticides, fungicides, nematicides, additional organic fertilizers, biopesticides, biological fungicides, biological nematicides, agricultural or horticultural adjuvants, stickers, spreaders, surfactants, dispersants, wetting agents, UV protectants, and mixtures thereof. The fertilizer may also contain organic carbon sources such as, but not limited to, compost and biochar.

Another aspect of the invention relates to a fertilizer comprising at least one microorganism selected from keratin degrading microorganisms and a keratin substrate acted on by the at least one microorganism to release nitrogen.

The method of the present aspect is carried out as in the previous aspect.

In one embodiment, the fertilizer comprises at least one microorganism selected from the group consisting of trichoderma viride (NRRL B-50520), scoparia glauca (NRRL B-50521), scoparia glauca (NRRL B-50522), and combinations thereof.

The keratin degrading microorganisms of the present aspect may include microorganisms of the genus Streptomyces (Streptomyces), including but not limited to Streptomyces avermitilis (Streptomyces avermitilis), Streptomyces coelicolor (Streptomyces coelicolor), and Streptomyces violaceus (Streptomyces violaceoruber).

This aspect of the invention may be carried out using any of the additives and any manner of application described above with respect to any of the above plants.

The method of the present aspect is carried out as in the previous aspect.

In one embodiment of this aspect of the invention, the fertilizer contains 5-20% by weight of gelatin based on the granules or pellets.

In practicing various aspects of the invention, the fertilizer may be prepared as a formulation containing organic or inorganic materials that facilitate the delivery or contact of the organism to the recipient plant or plant seed. Further, in all aspects of the invention described herein, the contacting of the fertilizer with the plant or plant seed or other plant material may be carried out simultaneously with, prior to, or subsequent to the introduction of the plant, plant seed or other plant propagation material into the growth medium or area. The plants, plant seeds, or other plant material may be formed (propagated) as described above, but not limited to, in any suitable growing medium and in any suitable environment (e.g., a greenhouse or field environment). The skilled person will be able to readily establish requirements suitable for maintaining and/or propagating a plant.

Regardless of the order in which the organisms are contacted with the plants, seeds or other plant material, the following are suitable methods for contacting the fertilizer with the selected plant material according to the invention. Non-limiting examples of such methods include broadcast application, drop application, spin application, liquid or dry furrow application, direct incorporation into soil or greenhouse planting mixtures, spray application, irrigation, injection, dusting, granulation, or coating a plant or plant seed or planting medium with a fertilizer. Granular formulations suitable for drop or broadcast application, powdered formulations added to potting mixtures or applied directly in the field, or liquid fertilizers using the systems described herein can also be produced.

The fertilizer of the present invention can be applied in the same manner as conventional fertilizers. Many methods and apparatus may be used, as known to those skilled in the relevant art. In one embodiment, the mixture of the microorganism of the invention and the substrate is applied directly to the soil or plant. In another embodiment, a dry powder of the microorganism of the invention with a substrate is applied to soil or plants. Fertilizer can be applied to the soil by spreaders, sprayers, and other mechanized devices that can be automated. The fertilizer may be applied directly to the plant, for example by soaking the seeds and/or roots, or spraying onto the leaves. Such administration may be performed periodically, such as once per year, or once per growing season, or more frequently as desired. Although not required, the fertilizer of the present invention may be used with other types of fertilizers or in rotation.

Beneficial microorganisms can be formulated or mixed to produce granules, powders, or liquid suspensions. These formulations can be mixed directly into the soil or planting mixture. The formulation is then mixed into The soil or into The volume of The planting mixture for greenhouse applications or into The upper volume of The field soil (Harman, G, "The Myths and Dogmas of biocontrol. changes in Perceptions Based on Research with Trichoderma harzianum T-22," Plant Disease 84: 377-. Equipment and procedures for such contact are well known and used in various agricultural industries. Typical rates are 10 per cubic meter of planting mixture or soil⁷To 10⁹0.2 to 10kg of product per colony forming unit (cfu).

Contact can also be made by liquid application (drench) for greenhouse or nursery soils and soil mixtures. Liquid suspensions of beneficial microorganisms can be prepared by: by mixing the dry powder formulation into water or other carrier, including fertilizer solutions, or by diluting the liquid formulation containing the organism in water or other aqueous solutions, including those containing fertilizers. In either case, the formulation may include other organic or inorganic additives to aid in dissolving or applying the mixture. Then, before or when plantingWhen the plant is actively growing, the solution can be used to sprinkle the planting mixture, such as by field irrigation. Usually, it contains 10⁷To 10⁹10 to 400ml of product (particle size typically 150/inn or less) of cfu is mixed with 100L of water for such applications.

Seeds are typically treated by methods well known in the industry using slurry, film coating or pelleting (Harman et al, "fans influencing Trichoderma hamatum Applied to Seeds As a Biocontrol Agent," Phytopathology 71: 569-. The microorganisms of the present invention may be effectively added to any such treatment provided that the formulation does not contain materials harmful to beneficial organisms. Depending on the organism in question, this may include chemical fungicides. Generally, a powder or liquid preparation of an organism (10)⁷To 10¹⁰cfu/g) are suspended in an aqueous suspending agent to obtain a biologically active level of the organism. The liquid typically contains a binder and other materials to provide a good level of coverage of the seeds and may also improve their planting shape or aesthetic appearance.

Contact may also be achieved by dry powders containing the beneficial organisms, which are applied to the roots, bulbs or seeds in powder form. Typically, a fine powder (typically 250/inch or less) is dusted onto the seeds, bulbs, or roots until the maximum load of powder is achieved (i.e., until no more powder will adhere to the treated surface). These powders generally contain 10⁷To 10⁹cfu/g。

Liquid suspensions of beneficial microorganisms can be prepared by: by mixing the dry powder formulation into water or other aqueous carrier, including fertilizer solutions, or by diluting a liquid formulation containing the microorganism in water or other aqueous solutions. These solutions can then be used to pour the planting mixture prior to planting or when the plant is actively growing. The liquid suspension of the product can be injected under pressure into the appropriate root zone of the plant through a hollow tube at the desired depth of application. For such applicationsAre well known in the horticultural industry. Alternatively, a suspension or powder containing the appropriate organism may be applied to the pores of the soil or other aqueous environment. Liquid suspensions of the product can be prepared as described above for the preparation of the infusion. Such materials may be added to furrows in which seeds are planted or plantlets are transplanted. Equipment for such applications is widely used in agriculture. Typical application rates are 0.5 to 10kg of product (10)⁷To 10⁹cfu/g)/hectare field.

The granules may be broadcast to the soil surface containing the growing plants, to the soil at the time of planting, or to the seeds or soil in which the plants are to be planted. Depending on the plant to be treated and the target of the treatment, a typical application rate range is 10⁷To 10⁹cfu/g of 1 to 500kg of product. Alternatively, Spray solutions may be prepared and applied to achieve similar rates (Harman, G.E., "The Dogmas and Myths of Biocontrol. changes in permissions Based on Research with Trichoderma harzianum T-22," Plant Dis.80: 736-.

For the purposes of the present invention, all methods describing application are designed to achieve the same objective, i.e., to provide an application method that will result in effective colonization of the root system by beneficial organisms (Harman et al, "patent and Existing Uses of Trichoderma and Gliocladium For Plant Disease Control and Plant Growth Enhancement," edited by Trichoderma and Gliocladium, Harman et al, Vol.2, London: Taylor and Francis (1998), which is incorporated herein by reference in its entirety).

The present invention can be used to treat seeds of a variety of plants. As used herein, the fertilizer of the present invention supports or enhances the growth of a plant, if the fertilizer in the soil is present, or applied to roots, stems, leaves, or other parts of the plant, the plant or other parts of the plant acquire viability, size, weight, germination rate, growth rate, or maturity rate. Thus, the fertilizer is suitable for use in any kind of agricultural, horticultural and forestry practice. Fertilizers can be used in large-scale commercial agriculture outdoors or in greenhouses, or even inside decorative plants. Preferably, the fertilizer is used to enhance the growth of crop plants, such as, but not limited to, cereal crops, vegetable crops, fruit crops, flower crops and grass crops. For example, the fertilizer composition may be used for dicotyledonous and monocotyledonous plants. More specifically, plants treated according to the present invention include any plant susceptible to infection by fungi or plant pathogens. For example, plants treated according to the present invention include, but are not limited to, agronomic intertillage or other field crops including buckwheat, legumes (soybean, taro, dried bean), corn (grain, seed, sweet corn, silage, popcorn, high oil corn), cotton, canola, peas (dry, juicy), peanuts, safflower, and sunflower; alfalfa hay and forage crops including alfalfa, clover, wild pea and clover; berries and small fruits including blackberry, blueberry, currant, elderberry, currant, blueberry, cranberry, raspberry, strawberry, grape, bulb crop: garlic, leeks, onions, green onions, and ornamental bulbs; citrus fruits including hybrid citrus, grapefruit, kumquat, lime, orange and grapefruit; melon vegetables including cucumbers, melons, gourds, pumpkins (pumpkins), pumpkins (squash), and fresh flowers; flower bed plants and ornamental plants; fruit vegetables including eggplant, sweet pepper, tomatillo, tomato, herb, spice, and mint; hydroponic crops including cucumbers, tomatoes and lettuce; herbs and spices; leafy vegetable and rape crops including arugula, celery, radishes, chicory, fennel, lettuce (head and leaves), parsley, chicory, rhubarb, spinach, swiss chard, cauliflower, brussels sprouts, cabbage, cauliflower, kale, headless cabbage, kohlrabi, mustard and asparagus; legume vegetables and field crops, which include tare and dried beans, lentils, juicy and dried peas, peanuts and soybeans; pome fruits including pears and sabina chinensis; crops including beetroot, beet, red beet, carrot, celeriac, chicory, horseradish, parsnip, radish kohlrabi, salsify, turnip; shadings and other nursery crops including deciduous trees (maple, oak), ornamentals, grapes, citrus, pine; small grain crops, including rye, wheat, sorghum, and millet; stone fruit, which includes apricot, cherry, nectarine, peach, plum, dried plum, woody nuts: almonds, beech nuts, brazil nuts, ash walnuts, cashews, chestnuts, hazelnuts, pecans, macadamia nuts, pecans, pistachios, and walnuts; tuber crops including potatoes, sweet potatoes, yams, jerusalem artichoke, cassava, and ginger. Other examples include those related to turf grass, turf, sports fields, parks, built and newly-built golf course tees, hole areas, fairways and rough areas, seed production, and turf production. The plants that can be treated also include petunia, geranium, poinsettia, chrysanthemum, carnation and zinnia.

The enhancement of plant growth of the present invention may take the form of larger root mass, greater rooting depth, larger shoot mass, larger shoot leaf length, increased leaf greenness, increased yield, and improved clumping and vigor. The growth of a plant can be established and determined by other means than the extrinsic properties listed above. One skilled in the art will be readily able to establish physical, biochemical or genetic tests to determine and/or quantify plant growth or viability.

Many advantages result from the fertilizer disclosed herein becoming colonized on root plants. One advantage is the prevention of Plant diseases by the direct Action of Microbial strains on Pathogenic microorganisms (see, for example, Chet, L, "Trichoderma-Application, Mode of Action, and Potential as a Biocontrol Agent of Soilborn Plant nutritional, in Innovative applications to Plant Disease Control, p. 137-160, Chet, J.Wiley and Sons: New York (1987), which is incorporated herein by reference in its entirety) or other harmful Soil microflora (Bakker et al," Microbial cell Production in the Rhizopheres in Retion to Point gravity Reduction and Pseudomonas-Mediated Plant Growth, "Soil Biochem.19: 451, which is incorporated herein by reference in its entirety). The microorganisms provide protection against plant pathogens due to the systemic induction of resistance by the resistance. This allows the Plant to be protected at points that are widely separated (either temporally or spatially) by the application of microorganisms (see Harman et al, "Trichoderma specifices-Opportunistic, Avirent Plant symbols," Nature Microbiol. Rev. 2: 43-56, (2004), which is incorporated herein by reference in its entirety). For example, by inducing resistance, the microorganisms disclosed herein can control foliar pathogens even when they are present only on the roots.

Another advantage is that the fertilizer of the invention can provide protection against abiotic stress due to drought (water deficit), disease or other adverse plant growth conditions. Many times, plants can be cultivated in climates where the crop is exposed to many biotic and abiotic stresses such as plant diseases and drought. Drought conditions affect gene expression, the amino acid normal profile and photosynthesis in plants to induce stress. In plants treated with the fertilizer of the invention, most of these reactions may be delayed. By treating plants with the microorganisms of the present invention in the field, it is possible to increase the plants' tolerance to drought. Plants with increased tolerance to drought, disease and stress would benefit farmers by stabilizing crop yield and profitability.

The microorganisms used in the disclosed fertilizers may result in larger and deeper roots and reduce the nitrogen demand for plant growth, presumably by increasing nitrogen uptake. This ability can also be used to reduce the nitrogen demand of plant producers. These strains may also improve the tolerance of plants to drought.

To understand the relevance of the present invention, it is important to examine the yield plateau of plants. Plants generally respond to increased nitrogen fertilizer levels with increased yield and growth to a point, then yield increases tend to plateau; this is the yield plateau beyond which the use of nitrogen fertilizers no longer increases yield. It has been shown that the microorganism of the present invention (i.e., Trichoderma viride (NRRL B-50)520) Broomcorn, scoparomyces brevis (NRRL B-50521) and scoparomyces brevis (NRRL B-50522)) treated planted seeds increased Plant growth and productivity even in the presence of significant nitrogen deficiency (see Harman, g., "Myths and Dogmas of biocontrol. changes in perfections from Research on Trichoderma harzianum T-22," Plant dis.84: 377-393 (2000); harman et al, "Enhancing Crop Performance and test Resistance with Genes from Biocontrol Fungi," in M.Vuro, J.Gressel, T.Butt, G.E.Harman, A.Pilgeram, R.J.St.Ledger and D.L.Nuss (eds.), Enhancing Biocontrol Agents and Handling Risks, pp.114-; harman et al, "Trichoderma specifices-Opportunistic, Avirule Plant sympositions," Nature Rev. Microbiol.2: 43-56(2004), which are hereby incorporated by reference in their entirety). Plants grown in the presence of symbiotic fungi are often greener and more viable (Harman, G., "Myths and Dogmas of biocontrol. changes in properties Derived from Research on Trichoderma harzianum T-22," Plant Dis.84: 377-393(2000), which is incorporated herein by reference in its entirety). Previous studies have found that yield plateaus are achieved with less than 40-50% nitrogen fertilizer in its presence compared to in the absence of Trichoderma Harman et al ("Enhancing Crop Performance and test Resistance with Genes from Biocontrol Fungi," in M.Vuro, J.Gressel, T.Butt, G.E.Harman, A.Pilger, R.J.St.Ledger and D.L.Nuss (eds.), Enhancing Biocontrol Agents and trading Risks, pp 114-. This means that the nitrogen fertilizer ratio of this amount can be reduced without a reduction in yield. This most likely reduces N evolution from the soil₂O, because the total fertilizer applied is reduced and a greater proportion of the applied nitrogen must be taken up by the plant, while the nitrogen requirement in the plant metabolism is not expected to change, therefore, the only way to obtain N addition in plants is through enhanced N use efficiency.

Another aspect of the invention relates to a method of making a fertilizer comprising an amino acid and ammonia. The method comprises providing at least one microorganism selected from the group consisting of: trichoderma viride (NRRL B-50520), Gliocladium brevicaulis (NRRL B-50521), Gliocladium brevicaulis (NRRL B-50522), and combinations thereof, and provides a substrate that is acted upon by at least one microorganism to release nitrogen. The method further comprises contacting the at least one microorganism with the substrate under conditions effective to cause the at least one microorganism to exert an action on the substrate and produce a liquid suspension comprising the amino acid and ammonia.

In this aspect of the invention, the fertilizer may be in liquefied form of all or part of the substrate (i.e., keratin) as a result of fermentation. Such fermentation will produce amino acids, peptides and/or ammonia.

Another aspect of the invention relates to an isolated Trichoderma viride strain deposited at the culture Collection of agricultural research services under accession number NRRL B-50520. The isolated Trichoderma viride strain may be in a biologically pure form.

Another aspect of the invention relates to an isolated strain of Scopulariopsis brevicaulis deposited at the agricultural research service culture Collection under the accession number NRRL B-50521. The isolated strain of Gliocladium brevicaulis may be in a biologically pure form.

Another aspect of the invention relates to an isolated strain of Scopulariopsis brevicaulis deposited at the agricultural research service culture Collection under the accession number NRRL B-50522. The isolated strain of Gliocladium brevicaulis may be in a biologically pure form.

Embodiment 1. a fertilizer comprising:

at least one microorganism selected from the group consisting of: trichoderma viride (NRRL B-50520), Scopulariopsis brevicaulis (NRRL B-50521), Scopulariopsis brevicaulis (NRRL B-50522) and combinations thereof, and

a substrate that acts to release nitrogen exerted by the at least one microorganism.

Embodiment 2. the fertilizer of embodiment 1, wherein the substrate is a protein selected from the group consisting of: feathers, feather meal and urea derivatives.

Embodiment 3. the fertilizer of embodiment 1, further comprising:

a complementing microorganism in the form of a trichoderma species.

Embodiment 4. the fertilizer of embodiment 3, wherein the supplemental microorganism is selected from the group consisting of: trichoderma viride, Trichoderma harzianum, Trichoderma atroviride, and combinations thereof.

Embodiment 5. the fertilizer of embodiment 1, further comprising:

a supplemental microorganism selected from the group consisting of: pyricularia indica, plant rhizosphere growth-promoting bacteria, mycorrhizal fungi and combinations thereof.

Embodiment 6. the fertilizer of embodiment 1, wherein the fertilizer is in the form of granules, pellets, dust, powder, slurry, film, and/or liquid suspension.

Embodiment 7. the fertilizer of embodiment 6, wherein the fertilizer is in the form of granules or pellets containing gelatin as a solidifying agent.

Embodiment 8 the fertilizer of embodiment 1, wherein the fertilizer comprises at least 10^4 colony forming units/g of the at least one microorganism in the substrate.

Embodiment 9. the fertilizer of embodiment 1, wherein the fertilizer has a nitrogen content of between 10-20 wt.%.

Embodiment 10 the fertilizer of embodiment 1, wherein the fertilizer further comprises:

a non-protein binding agent selected from the group consisting of dextran, starch, polyvinyl chloride, and combinations thereof.

Embodiment 11 the fertilizer of embodiment 1, wherein the fertilizer further comprises:

a soluble nitrogen source selected from the group consisting of nitrate, ammonia, ammonium salts, amino acids, urea, fish meal or extract, compost extract, seaweed extract, shrimp extract, shellfish extract, and combinations thereof.

Embodiment 12. the fertilizer of embodiment 1, wherein the fertilizer further comprises:

a source of phosphorus selected from the group consisting of rock phosphate, sodium phosphate, potassium phosphate, bone meal, and combinations thereof.

Embodiment 13. the fertilizer of embodiment 1, wherein the fertilizer further comprises:

a potassium source selected from the group consisting of potassium chloride, potassium phosphate, potassium sulfate, green sand of New Jersey, an organic source of potassium, and combinations thereof.

Embodiment 14. the fertilizer of embodiment 1, wherein the fertilizer further comprises:

a micronutrient selected from the group consisting of salt, substances containing iron, cobalt, manganese, magnesium, copper, calcium, boron, zinc, and combinations thereof.

Embodiment 15 the fertilizer of embodiment 1, wherein the fertilizer further comprises:

and (5) growing a medium.

Embodiment 16 the fertilizer of embodiment 15, wherein the growth medium is selected from the group consisting of: soil, sand compost, peat, rice hulls, coconut fibre, coconut coir, soilless growth media containing organic and/or inorganic components, artificial plant growth media, polymer-based growth media, hydroponic nutrients and growth solutions, organic soil amendments, water and mixtures thereof.

Embodiment 17. the fertilizer of embodiment 1, wherein the fertilizer further comprises:

a carrier selected from the group consisting of water, aqueous solutions, slurries, and powders.

Embodiment 18 the fertilizer of embodiment 1, wherein the fertilizer further comprises:

an additive selected from the group consisting of: insecticides, fungicides, nematicides, additional organic fertilizers, biopesticides, biological fungicides, biological nematicides, agricultural or horticultural adjuvants, adhesives, spreaders, surfactants, dispersants, wetting agents, UV protectors and mixtures thereof.

Embodiment 19 the fertilizer of embodiment 1, wherein the fertilizer further comprises:

an organic carbon source selected from the group consisting of compost and biochar.

Embodiment 20. a fertilizer comprising:

at least one microorganism selected from keratin-degrading microorganisms, and

a keratin substrate that is acted upon by the at least one microorganism to release nitrogen.

Embodiment 21 the fertilizer of embodiment 20, wherein the at least one microorganism is selected from the group consisting of trichoderma viride (NRRL B-50520), scoparia glauca (NRRL B-50521), scoparia glauca (NRRL B-50522), and combinations thereof.

Embodiment 22 the fertilizer of embodiment 20, further comprising:

at least one microorganism selected from the group consisting of Streptomyces avermitilis, Streptomyces coelicolor, Streptomyces violaceus, and combinations thereof.

Embodiment 23. the fertilizer of embodiment 20, further comprising:

a complementing microorganism in the form of a trichoderma species.

Embodiment 24 the fertilizer of embodiment 23, wherein the supplemental microorganism is selected from the group consisting of: trichoderma viride, Trichoderma harzianum, and Trichoderma atroviride, and combinations thereof.

Embodiment 25. the fertilizer of embodiment 20, further comprising:

Embodiment 26 the fertilizer of embodiment 20, wherein the fertilizer is in the form of granules, pellets, dust, powder, slurry, film, and/or liquid suspension.

Embodiment 27 the fertilizer of embodiment 20, wherein the fertilizer is in the form of a liquid suspension comprising an amino acid and ammonia.

Embodiment 28. a method of enhancing plant growth, the method comprising:

providing a fertilizer comprising:

a substrate that is acted upon by the at least one microorganism to release nitrogen; and is

Contacting the fertilizer with the plant or plant seed under conditions effective to enhance the growth of the plant or plant seed compared to the plant or plant seed without the application of the fertilizer.

Embodiment 29 the method of embodiment 28, wherein the substrate is a protein selected from the group consisting of: feathers, feather meal and urea derivatives.

Embodiment 30. the method of embodiment 28, further comprising:

a complementing microorganism in the form of a trichoderma species.

Embodiment 31 the fertilizer of embodiment 30, wherein the supplemental microorganism is selected from the group consisting of: trichoderma viride, Trichoderma harzianum, and Trichoderma atroviride, and combinations thereof.

Embodiment 32 the method of embodiment 28, further comprising:

Embodiment 33 the method of embodiment 28, wherein the fertilizer is in the form of granules, pellets, dust, powder, slurry, film, and/or liquid suspension.

Embodiment 34 the method of embodiment 33, wherein the fertilizer is in the form of granules or pellets containing gelatin as a solidifying agent.

Embodiment 35 the method of embodiment 34, wherein the gelatin comprises 5-20% by weight of the granules or pellets.

Embodiment 36 the fertilizer of embodiment 33, wherein the fertilizer is in the form of a liquid suspension comprising an amino acid and ammonia.

Embodiment 37 the method of embodiment 28, wherein the fertilizer has a nitrogen content of between 10-20 wt.%.

Embodiment 38 the method of embodiment 28, wherein the fertilizer further comprises:

Embodiment 39 the method of embodiment 28, wherein the fertilizer further comprises:

a soluble nitrogen source selected from the group consisting of nitrates, inorganic ammonia sources, ammonia, ammonium salts, amino acids, urea, fish meal or extract, compost extract, seaweed extract, shrimp extract, shellfish extract, and combinations thereof.

Embodiment 40 the method of embodiment 28, wherein the fertilizer further comprises:

Embodiment 41 the method of embodiment 28, wherein the fertilizer further comprises:

Embodiment 42 the method of embodiment 28, wherein the fertilizer further comprises:

a micronutrient selected from the group consisting of salt, other substances containing iron, cobalt, manganese, magnesium, copper, calcium, boron, zinc, and combinations thereof.

Embodiment 43 the method of embodiment 28, wherein the fertilizer further comprises:

and (5) growing a medium.

Embodiment 44 the method of embodiment 43, wherein said contacting is performed before said plant or said plant seed is in said growth medium.

Embodiment 45 the method of embodiment 43, wherein said contacting is performed after said plant or said plant seed is in said growth medium.

Embodiment 46. the method of embodiment 43, wherein the growth medium is selected from the group consisting of: soil, sand compost, peat, rice hulls, coconut fibre, coconut coir, soilless growth media containing organic and/or inorganic components, artificial plant growth media, polymer-based growth media, hydroponic nutrients and growth solutions, organic soil amendments, water and mixtures thereof.

Embodiment 47 the method of embodiment 28, wherein the fertilizer further comprises:

Embodiment 48 the method of embodiment 28, wherein the fertilizer further comprises:

an additive selected from the group consisting of: insecticides, fungicides, nematicides, additional organic fertilizers, biopesticides, biological fungicides, biological nematicides, agricultural or horticultural adjuvants, stickers, spreaders, surfactants, dispersants, wetting agents and UV protectants and mixtures thereof.

Embodiment 49 the method of embodiment 28, wherein the fertilizer further comprises:

Embodiment 50 the method of embodiment 28, wherein the plant or plant seed is selected from the group consisting of: buckwheat, beans, corn, cotton, canola, peas, peanuts, safflower, sunflower, alfalfa, clover, vetch, axanthus, blackberry, blueberry, gooseberry, elderberry, gooseberry, huckleberry, loganberry, strawberry, grape, garlic, leek, onion, shallot, ornamental corm, citrus hybrids, grapefruit, kumquat, lime, orange, grapefruit, cucumber, melon, cucurbit, pumpkin, squash, fresh flowers, altar and ornamental plants, eggplant, sweet pepper, mucuna puree, tomato, herbs, spices, mint, sesames, celery, radish, chicory, fennel, lettuce, parsley, chicory, rhubarb, spinach, lettuce, cauliflower, brussel sprout, cabbage, cauliflower, kale, kohlrabi, mustard, asparagus, lentils, pear, quince, beet root, beet, red beet, etc, Carrot, celeriac, chicory, horseradish, parsnip, radish turnip, salsify, turnip, deciduous tree, ornamental plants, citrus, pine, rye, wheat, sorghum, millet, apricot, cherry, nectarine, peach, plum, dried plum, almond, beech nut, brazil nut, juglans regia, cashew nut, chestnut, hazelnut, pecan nut, macadamia nut, pecan nut, pistachio nut, walnut, potato, sweet potato, yam, jerusalem artichoke, cassava, ginger, and turfgrass.

Embodiment 51. the method of embodiment 28, wherein the contacting is performed by: broadcast application, drop application, rotary application, liquid or dry in-furrow application, direct incorporation into soil or greenhouse planting mixtures, spray application, irrigation, injection, dusting, extrusion, granulation, or coating the plant or the plant seed with the fertilizer.

Embodiment 52. the method of embodiment 28, wherein the plant growth enhancement is in the form of a larger root mass, a larger rooting depth, a larger shoot mass, a larger shoot leaf length, increased leaf greenness, increased yield, and improved clumping and vigor.

Embodiment 53 a method of making a fertilizer comprising an amino acid and ammonia, the method comprising:

providing at least one microorganism selected from the group consisting of: trichoderma viride (NRRL B-50520), Gliocladium brevicaulis (NRRL B-50521), Gliocladium brevicaulis (NRRL B-50522), and combinations thereof;

providing a substrate that is acted upon by the at least one microorganism to release nitrogen; and is

Contacting the at least one microorganism with the substrate under conditions effective to cause the at least one microorganism to exert an action on the substrate and produce a liquid suspension comprising an amino acid and ammonia.

Embodiment 54. a fertilizer produced by the method of embodiment 53.

Embodiment 55, an isolated strain of trichoderma viride deposited with the culture collection of the agricultural research service under accession number NRRL B-50520.

Embodiment 56. the isolated trichoderma viride strain of embodiment 55, which is biologically pure.

Embodiment 57, an isolated strain of scopulariopsis brevicaulis deposited at the agricultural research service culture collection under accession number NRRL B-50521.

Embodiment 58. the isolated strain of scopulariaceae brevicaulis as described in embodiment 57, which is biologically pure.

Embodiment 59. an isolated strain of scopulariopsis brevicaulis deposited at the agricultural research services culture collection under accession number NRRL B-50522.

Embodiment 60. the isolated strain of scopulariaceae brevicaulis as described in embodiment 59, which is biologically pure.

The following examples are provided to illustrate embodiments of the present invention and are in no way intended to limit the scope thereof.

Example 1 isolation of efficient microorganisms for degradation of waste proteins

One of the most widespread proteins found in agricultural waste is keratin. This is a structural component of hair, feathers and other similar materials. Although keratin generally degrades slowly in soil or elsewhere, there are also microorganisms that degrade this substrate relatively quickly.

The organisms of the invention can grow in liquid fermentation and will release ammonia and amino acids. For the use of the strain NRRL described herein, additional nutrients may be used, such as trypticase soy broth (product of Difco). 1 to 10% w/v feather meal is added to water containing trypticase soy broth and the mixture is sterilized by autoclaving. When organisms are added to this sterile medium, the feather meal is degraded. The ammonia is released from the feather meal creating an alkaline environment. Since ammonia at high pH is strongly volatile, the presence of this material is easily detected by odor. Systems for both harvesting and continuous removal of ammonia are readily available and known to those skilled in the art. One approach is to simply place a closed sterile closed loop outside of the fermentation vessel that includes a sterile air injection device. When air is introduced through the fermentation broth, ammonia will be removed and can be trapped by transfer into the acidic solution, which vinegar will be a satisfactory acidic solution. This will result in the formation of ammonium acetate which can be readily used as a fertilizer, and this sparging step will result in an enrichment of ammonia for further use. If removal of amino acids is also required, this step can be carried out by passing the fermentation broth through a suitable ion exchange resin from which the amino acids are eluted and thereby enriched. At the same time, the sparging process will remove excess ammonia that raises the pH to allow fermentation to continue. This can be developed as a continuous flow system for efficient ammonia production.

The present invention seeks to identify effective microorganisms for degrading proteins. To achieve this, feather powder was mixed with peat moss and the mixture was wetted (composition: 1L peat moss, 1L feather powder and 400ml water), autoclaved for two consecutive days and inoculated with 1g of composted chicken manure. Peat moss is used as an accretion/aeration component and composted chicken manure is considered a good source of keratin (i.e. feather) degrading microorganisms. Within six days of inoculation, there was a strong ammonia smell, indicating that the feathers degraded into ammonia, which is an excellent natural nutrient for plants. Organisms present in the ammonia odorous medium were plated on potato dextrose agar ("PDA") ("Difco") and trypticase soy agar ("TSA") ("Difco") prepared according to the manufacturer's instructions using a dilution technique. Microbial growth on PDA, which is typically a fungal medium, is generally poor, but on TSA, which is hydrolyzed soy protein, many fungi grow. Surprisingly, these microorganisms consist of only three different microorganisms which apparently grow well on the feather meal, while the other most abundant microorganisms present in the compost do not grow well on the feather meal. Isolation by single spores, obtaining pure forms of these fungi and storing on silica gel at-20 ℃ is a very good way of preserving the fungi.

Fungi isolated from chicken manure compost were inoculated into trypticase casein soy broth ("TSB") and grown for three days. Good growth of all microorganisms was obtained in TSB and 50ml of actively growing culture was added to feather meal contained in a sterile plastic box with a flange lid: in peat mixtures, the lid provides good aeration of the contents of the box. Over a six day growth, samples were removed, extracted and the level of soluble nitrogen compounds determined. The extracts were analyzed for amino acids (ninhydrin method) or ammonia (Nessler reagent; LaMotte test kit).

Promising results were obtained with the obtained fungi. The ammonia level of the control sample was about 19. mu.g/ml based on lysine, while the values for the three strains were 600. mu.g/g, 3240. mu.g/g and 4900. mu.g/g, respectively. The amino acid levels of the same samples were 2800. mu.g/g, 3003. mu.g/g and 5600. mu.g/g for the same three strains, while the control value was about 3. mu.g/ml. These measurements were repeated several times and the results were similar. In particular, several strains are more effective in degrading keratin than any other tested microorganism.

Example 2 identification of Keratin-degrading fungi and bacteria

The fungi identified in example 1 above were sent to the USDA type culture collection for deposit in their patent collection. These strains were designated NRRL B-50520, NRRL B-50521, and NRRL B-50522 for the green, brown, and white spore strains, respectively. These fungi were identified as Trichoderma viride (NRRL B-50520) and Gliocladium brevicaulis (NRRL B-50521 and NRRL B-50522 are two isolates of the fungus). These two species have brookform conidia, but the subtle differences in the arrangement of the conidia clearly place them in the respective genera. The ITS regions of each isolate were also sequenced and evaluated by GenBank BLAST queries. As a result, their identities are confirmed as follows:

NRRL B-50520-Trichoderma viride

NRRL B-50521-Scopulariopsis brevicaulis

NRRL B-50522-Scopulariopsis brevicaulis

Trichoderma viride has a very low risk profile. The U.S. environmental protection agency states that "the agency predicts that the current registration sites for trichoderma polyspora and trichoderma viride do not require additional health-impact data relative to the use of strains in this genus as biocontrol agents for fungal pathogens" (trichoderma species profile, registration review: original case, U.S. environmental protection agency (2007 month 4), which is incorporated herein by reference in its entirety). The same applies to most other fungi of the genus Trichoderma.

Recent reviews list keratinase-producing organisms (Brandelli et al, "biological feedstocks of Microbial keratanases and Their Production and Applications," Applied Microbiology and Biotechnology 85: 1735-1750(2010), which is hereby incorporated by reference in its entirety). Gliocladium brevicaulis is described as a producer of keratinase, but Trichoderma viride is not. However, there is a single report From India for the production of keratinase by Trichoderma viride strains (Jyoti et al, "Keratinolistic Enzymes From Trichoderma viride and Graphium meneianum Isolated From Poultry farm at Jabalpur, India," Cryptogamic Botany 4: 30-33(1993), which is incorporated herein by reference in its entirety).

Example 3 method for producing useful formulations

Feather meal is a coarse brown powder. Dry fertilizers used in many applications need to be applied in granular form by dropping or rotating dispersion systems. Thus, processes for preparing useful particles have been developed. A method of solidifying and making particles having a diameter size of 2-5 mm is required. The starting particles were prepared by: sufficient water was added to the feather powder to make a slurry, and then gelatin was dissolved in the slurry to a final concentration of about 7% relative to the feather powder. This material was dispersed in the form of flakes and dried, resulting in hard brown flakes. This was subsequently ground and sieved to obtain particles of the desired size. It will be appreciated that similar compositions may be extruded and dried to give suitably sized particles. Similarly, these mixtures can also be used in granulation equipment, subsequently appropriately sized.

The resulting granules do not contain any added microorganisms. Some systems for making granules or pellets involve heating and applying pressure, and therefore coating procedures for pellets have been developed. Conidia of a trichoderma strain known to provide benefits, including enhanced nitrogen use efficiency, were added to 0.1% Crystal Tex (cassava dextran, National Starch) to yield about 10⁵Individual colony forming units/ml of suspension. This mixture is applied to the surface of the dried pellets to give a smooth and continuous coating that does not significantly increase the size of the particles. Thus, the live fungus is applied as a dry coating. From this position, these fungi immediately grow when the pellets are wet (fig. 1).

In later experiments, the strains were added directly to the mixture while wet and then extruded to form pellets for inspection. This method works very well and the organism remains stable as long as the drying temperature is kept at 38 ℃ or below. The selection of the appropriate microorganism (i.e., microorganism) helps to achieve the best results. These microorganisms have two functions: (1) certain microorganisms have the ability to degrade feather meal or other resistant proteins, and (2) certain microorganisms act as endophyte symbiota. In the experiments of the present invention, N, as judged by green turning of lawn plants, occurred only 4-6 weeks after application in the absence of microorganisms. At this point, most of the N has been released in the presence of degrading microorganisms. Endophytic plant microorganisms have many functions, one of which is to increase the nitrogen utilization efficiency ("NUE") of plants. If this microbial component is included, the effective N value of the fertilizer increases. Thus, when the NUE contribution is included, the amount of N in the 14% N fertilizer actually becomes 28%. This is a useful aspect of the invention if it competes with and is as effective as a standard synthetic fertilizer having 30% N. Plants still require the same level of N to thrive, so the only way for the fertilizer of the present invention to become equally effective (useful for economic competitiveness) at the same application rate is to enhance the organism by using NUE.

Standard synthetic fertilizers contain high levels of salt and the osmotic potential and direct effects of these components are lethal to microorganisms. However, in the present invention, the synthetic salt is almost completely eliminated, and therefore, the microorganism-containing particles are safe for microorganisms. Most particles are proteins, which are non-toxic to microorganisms. Therefore, the application or direct mixing of the microbial agent is one of the constituents of the present invention.

The microbial component and the non-biological component may be formulated as a single granule or other mixture. There are many of the following situations in the literature: among these, for example, microorganisms added to or living in soil increase plant growth, including NUE. However, users are reluctant to administer multiple different organisms and other products to achieve valuable results. It is an aspect of the present invention to incorporate all biological and non-biological components into a single product or granule.

This embodiment of the invention provides particles that are almost entirely composed of protein. The protein is 12-14% N, so the necessary values were obtained because both gelatin and feather meal are animal proteins.

For some applications, a granular product is not necessary for some applications. Powders are often used as components of, for example, plant growth media, and as materials for direct addition to the field or garden, typical nitrogen values for particles such as these are 12-14%. The mixture of feather meal and microorganisms described herein may be used directly.

In addition, any number of additives may be made into such primary particles or powders. One example is a liquid ammonia and amino acid mixture extracted from the growth medium as in example 2 above. This can be used directly as a dry powder or as granules, or concentrated to give higher levels of N. Such addition can be used to provide immediate greenness return since feather meal or gelatin provide little N immediately available. Other suitable nitrogen sources are fish meal or extracts, compost extracts, seaweed extracts, marine animals such as shrimp or shellfish extracts. Inorganic sources include nitrates, inorganic ammonia sources, and the like. In addition, it is possible to add the products listed above, such as K from green sand or potassium sulfate listed above, and other K sources not listed. Similarly, P may be provided in bone meal (organic) form or from various inorganic salts. For lawn applications, P is prohibited and the use of feather meal or gelatin components is because P is absent.

Example 4 Fertilizer for grass

A very demanding use of fertilizers is application to grass in lawns, sports fields, parks, golf courses and other similar applications. The fertilizer must provide an extremely attractive surface and last for a considerable period of time. Granules were prepared to contain 73.8% feather meal, 13% chilium nitrate, 7.3% gelatin, and 5.7% new jersey greensand (K source). Chilium nitrate is currently organically certified and contains 16% N, so its nitrogen content is only slightly higher than that of the protein component. As a comparison, the formulation produced relative to the assay was 30: 2: 8N: P: K (at N, P)₂O₅And K₂O form) widely used household fertilizer

To perform the test. By comparison, it is expected that the analysis of the fertilizer prepared is 15-16: 0: 0.3, so it is not a complete fertilizer. P is not included because of the risk of contaminating the water body with phosphorus resulting in eutrophication, which is not allowed in many jurisdictions. Ratio of N levels

The N level of the fertilizer is as low as 50%, and the K level is suboptimal. Thus, if the performance of the fertilizer formulation of the present invention is close to that of the fertilizer formulation

Which illustrates that (a) significant amounts of N are released from the fertilizer, (b) increased N uptake efficiency is induced in the lawn plants by the added fungus, and (c) less available N permeates into groundwater. After all, plants also require the same amount of N to grow, and if only half of the nitrogen is added, then there must be greater N uptake and only a small amount of nitrogen that penetrates into the groundwater.

In 1: containing little free nitrogen1 sandy loam soil: tests were performed on annual ryegrass planted in peat mixtures. Annual ryegrass is planted in a shallow seedbed and various fertilizers are applied after about two weeks after planting after seedlings have formed. The granules were applied at a rate of 1x, this being so

The recommended application rate to the level of fertilizer applied. In addition, each treatment was applied twice at this rate. The first experiment included some of the following treatments:

no treatment control

Administration of particles without any microorganisms

Particles with B-50520 applied

Particles coated with B-50521

Particles coated with B-50522

Particles coated with B-50520+ Trichoderma strain K1-K4 (described in the next example)

Particles coated with B-50521+ Trichoderma strain K1-K4

Particles coated with B-50522+ Trichoderma strain K1-K4.

·

Fertilizer

All were applied at 1x and 2x application rates. In this example, K1-K4 consisted of the following strains: trichoderma harzianum strain RR17Bc (ATCC accession PTA 9708); trichoderma harzianum F11Bab (ATCC accession No. PTA 9709), Trichoderma atroviride strain WW10TC4(ATCC accession No. 9707), and Trichoderma viride strain 41(ATCC accession No. 20476).

The results of these tests were successful and are summarized in FIGS. 2A-2B and FIGS. 3A-3D.

In FIG. 2A, the treatments used were PT (organic fertilizer, NRRL B-50521, and K1-K4, which include Trichoderma harzianum strain RR17Bc (ATCC accession PTA 9708), Trichoderma harzianum F11Bab (ATCC accession PTA 9709), Trichoderma atroviride strain WW10TC4(ATCCPTA accession 9707), and Trichoderma viride strain 41(ATCC accession 20476));

p ═ organic fertilizer with NRRL B-50521; and C ═ untreated control. In FIG. 2B, the treatments used were PT (organic fertilizer and NRRL B-50521); OFC ═ organic fertilizer granules without any fungi; p (organic fertilizer with NRRL B-50521). Fig. 2A-2B show growth approximately 10 days after fertilization by dispersing the particles on the surface of the soil where the grass is growing. The growth increase is mainly caused by available soluble nitrogen. As can be seen, the growth of grass using the organic fertilizer was substantially greater in the presence of NRRL B-50521 as compared to any treatment without such organisms. Similar results are seen with either NRRL B-50520 or NRRL B-50522. During this time period, grass growth was also greater compared to commercial lawn fertilizer. The organic fertilizer control also grew better than the control without granules; this can be partly attributed to the small amount of organic chilium nitrate (mainly NaNO) added to the granules as starting fertilizer₃). This illustrates the utility of the invention disclosed herein including microorganism-mediated slow release of nitrogen.

Fig. 3A-3D contain graphs showing growth over time. To obtain these data, grass was cut from a shallow seedbed, dried and weighed to simulate a mowing event (in fig. 2A-2B, grass was prepared for cutting), and this continued throughout the experiment. All cuts were made on the same day, so the figures represent the amount of grass harvested between the last cut and the next cut, and the days after harvesting represent the days after fertilization when the cuts were made.

On day 7 after sowing, the untreated grass, at its highest cut level, was 800 mg/shallow bed, after which the cut weight dropped to a very low level (fig. 3A). The organic fertilizer at 1x application rate was similar to the untreated control, but at 2x application rate, the weight was significantly greater. The organic fertilizer was higher at the first cut, which undoubtedly represents the nitrogen provided by the chilium nitrate provided in the granules. After one week, the grass weight decreased and then increased again (fig. 3A). This second increase is undoubtedly due to the hydrolysis of the feather meal by natural soil microflora.

Standard of merit

Moderate growth was initially supported at the 1x level, but this growth increased upon the second cut. Growth declined again at the third cut and finally increased again at the last cut (fig. 3B). This may reflect a change in the availability of different sources of nitrogen. However, at double application rate, grass yield was very low at the second and third cut (fig. 3B). This is caused by grass burn at this application level, which never occurred in any feather meal based system.

At 1x application rate, strain NRRL B-50520 produced grass cutting yield results that were significantly better than the control organic fertilizer (fig. 3A and 3C). Strain NRRL B-50521 produced a rapid release of nitrogen as evidenced by a very high grass yield on the first cut, but then in most cases the grass yield decreased greatly on the second cut. This is due to the very high soluble nitrogen release followed by substrate depletion.

In these experiments, it should be noted that,

the fertilizer was relatively complete, while, as previously mentioned, the test material used contained no phosphorus and insufficient levels of potassium.

Example 5 library size of soluble Nitrogen Compounds in soils and grasses fertilized with different materials

It is shown in the above examples that fertilizers prepared by other methods will have low levels of nitrate leaching into groundwater. If this is true, the nitrate levels should be lower than in soil fertilized with standard chemical fertilizers. Also, other forms of soluble nitrogen should be increased due to the degradation of feather meal resulting in the release of amino acids and ammonia. These compounds will be converted over time to nitrate only by the activity of the soil microflora. However, it is likely that amino acids and ammonia will be rapidly absorbed by plants (or microflora) and thus the pool size will not accumulate to large levels. Thus, the amount of free nitrogen in any form may be smaller in the soil with the sample of the present invention compared to the soil with the standard chemical fertilizer. Data from soil sampled nine days after fertilization are listed in table 1 below. Every 3.6ml extraction buffer 1g of soil was extracted and the colorimetric test described above was used to measure soluble nitrogen in the extract. With respect to the levels of amino acids, the data are based on dilution curves with lysine as standard. All these determinations were performed at 1x fertilization rate.

TABLE 1Amount of soluble nitrogen in different forms in the soil

Nitrate values in the soil were in full agreement with the predictions. Using a chemical standard fertilizer, the level of nitrate was an order of magnitude greater than that of the control soil with any feather meal based granular product without treatment. The ammonia levels did not vary much between treatments, probably because any ammonia released was immediately absorbed by the already grown and formed turf. However, the level of amino acids changes. The maximum level of amino acids was granules with NRRL B-50521. This correlates well with a rapid increase in turf growth when fertilized with particles containing this organism at an early stage (fig. 3D). Interestingly, the addition of K1-K4 to particles containing either B-50520 or B-50521 reduced the amino acid level. This may be caused by significant conversion of the reduced form of nitrogen (amino acids or ammonia) to nitrate. If the concept of enhancing turf growth by in vivo hydrolysis of feather meal is correct, the size of the nitrogenous compound pool in the turf plant itself should vary in quantity and quality. Table 2 gives the concentrations of nitrate, ammonia, amino acids (as lysine equivalents) in plant tissue extracted from the grass seedlings 14 days after planting. In these experiments, the grass was dried and then extracted, and the levels of the various soluble nitrogen forms were measured by the procedure already described.

TABLE 2The concentration of nitrate, ammonia and amino acids in plant tissue

This data confirms the following expectation: the source of nitrogen is such that the soluble nitrogen form in plant tissues is different. It must be emphasized that these values are snapshots of the size of the active nitrogen metabolism pool in plants and that there is a constant rebalancing between uptake from the environment and the metabolism of nitrogen in turf into proteins and other nitrogen-containing compounds. Untreated controls had low levels of various forms of nitrogen, which explains the poor growth noted above. The ammonia levels varied over a small range except for B-50520 and B-50521 on the granules, but in the case of both fungi on the granules, the levels were rather high. If K1-K4 is added to B-50520 or B-50521, the level of ammonia appears to decrease. This can be illustrated by the conversion of ammonia to nitrate by these fungi, as suggested by the increased nitrate in seedlings grown in the presence of both fungi.

Seedlings grown in the presence of commercial fertilizers had very high levels of nitrate, again as expected. The main nitrogen-containing component of this fertilizer is nitrate and this is clearly reflected in the values obtained. High levels of nitrate are contaminants of water and these data reflect the points at which the following occurs: the amount of nitrate saturates the plant's ability to absorb this nutrient and most of this nutrient will end up as water and air pollutants. Seedlings grown in the presence of the feather meal particles and microbial mixture K1-K4, and NRRL B-50520 or B-50521 have only about 2% of the nitrate in their tissues as those grown in the presence of inorganic lawn fertilizer. As seen in the previous examples, their growth is much better than predicted by nitrate levels in their tissues. However, as with the level of ammonia, the pool size of amino acids is enhanced. In plant assimilation and nitrogen conversion, amino acids and ammonia are downstream of nitrate-in plants, nitrate needs to be converted to ammonia and then to amino acids before it can be absorbed and converted to a structure or active ingredient metabolized by the plant. Therefore, this higher level of available N would improve plant growth and nitrogen utilization efficiency without the energy-wasting step of reducing to ammonia. The possibility of greatly reducing nitrate contamination for ground and surface waters is also demonstrated. Surprisingly, nitrate levels in plant tissues increased when either B-50520 or B-50521 was added with K1-K4. This may be due to interactions between these groups of fungi that result in greater oxidation of amino acids or ammonia to nitrate.

Example 6 production of Biopreferred Fertilizer preparation

As shown in FIG. 1, the recipes for the products used to generate the data in this figure are roughly the same as

The fertilizer supported the growth of the grass equally well and only at slightly elevated application rates for about 5 weeks. However, after that, the results are less effective, and

the synthetic fertilizer was effective for approximately 8 weeks.

Furthermore, the price of feather meal was found to increase 2-3 times in the summer of 2012, which exceeds the price of the product containing it. The answer appears to be the production of bioprefred, not a product that can be certified by OMRI. As mentioned before, the bioperfred product must contain minimally 71% of the total organic farm material. Thus, synthetic slow-release nitrogen sources with higher N levels can be used. Granules were prepared containing 71% organic protein (mainly feather meal), 20% methylene urea (slow release synthetic nitrogen source) by dry weight, the remainder being divided between potassium sulfate and chilium nitrate. The N to P to K ratio is about 17 to 0 to 9, and the slow release of N is expected to provide an extended life of the product. The granules so produced are coated or otherwise contain Trichoderma strain K1-K4 plus Trichoderma viride NRRL B-50520 to provide NUE benefits to product properties equivalent to those with 32% N

In addition, another product was prepared. The product contains 46% of organic protein source, mainly feather meal and 26% garden compost. It also included methylene urea at a level of 23% of the total, with the remainder consisting of chilium nitrate and potassium sulfate. The product had an N: P: K ratio of 16: 0: 3 and contained 71% of the Biopreferred fraction. This product is expected to have similar capabilities as the product described in the preceding paragraph; however, due to the reduced amount of organic proteins, it is expected to be about 20% cheaper. It is also an advantage to have a greater amount of organic material from the compost in the formulation.

The granules may also contain a non-protein binder (e.g., gelatin), such as dextran, starch, polyvinyl chloride, or other similar materials.

Particles produced using the techniques and systems described herein perform well in actual testing. However, when combined (whether by coating the particles with beneficial microorganisms or internally incorporated), they provide their best performance. FIG. 4 shows an example of increased development of a root system of a lawn. Strains K1-K4 have repeatedly been shown to increase plant root growth by seed treatment or other methods applied in wheat, corn, tomato and other crops. This property that increases plant root development and deeper colonization of the soil is one component of the ability of K1-K4.

Fig. 4 shows the results in sand: turf grown for about 6 weeks in peat mixtures and fertilized with the granules described in this example with and without the beneficial microbial mixture trichoderma viride strain NRRL B-50521 and trichoderma strain K1-K4 developed roots. Roots grown on particle-fertilized turf without beneficial microorganisms are shown in petri dishes labeled 11. The petri dish labeled 12 contains a medium of about 10⁵cfu/g organisms incorporated in the particles, while the 13 marked petri dish was fertilized with particles of beneficial fungi coated in a glucan mixture (10 in 2% tapioca starch glucan)⁵Colony forming units (cfu)/ml to form one continuous thin coating). These are methods of adding beneficial microorganismsA change in (c). Greater root growth (turf roots are rather small and delicate) is evident in the lower half of the petri dish using either method of application of beneficial microorganisms, but is absent in turf fertilized with particles in which beneficial microorganisms are not present.

The present invention has two microbial components. First, microorganisms that increase the degradation of naturally resistant N sources, such as feather meal, to convert them into simple compounds that can be used for plant nutrition. Second, microorganisms having the ability to increase plant growth and productivity, including enhancing root growth, NUE, increasing tolerance to abiotic stress, and increasing the photosynthetic capacity of the plant, such as K1-K4.

Although preferred embodiments have been depicted and described in detail herein, it will be apparent to those skilled in the relevant art that various modifications, additions, substitutions and the like can be made without departing from the spirit of the invention and these are therefore considered to be within the scope of the invention as defined in the following claims.

Claims

1. A fertilizer, comprising:

trichoderma viride NRRL B-50520 microorganism, and

a protein substrate that acts to release nitrogen exerted by the microorganism.

2. The fertilizer of claim 1, wherein the protein substrate is selected from the group consisting of: feathers and feather meal.

3. The fertilizer of claim 1, further comprising:

a complementing microorganism in the form of a trichoderma species.

4. The fertilizer of claim 3, wherein the supplemental microorganism is selected from the group consisting of: trichoderma viride, Trichoderma harzianum, Trichoderma atroviride, and combinations thereof.

5. The fertilizer of claim 1, further comprising:

6. The fertilizer of claim 1, wherein the fertilizer is in the form of granules, powder, slurry, film, and/or liquid suspension.

7. The fertilizer of claim 6, wherein the fertilizer is in the form of granules containing gelatin as a solidifying agent.

8. The fertilizer of claim 1, wherein said fertilizer comprises at least 10^4 colony forming units/g of said microorganism in said protein substrate.

9. The fertilizer of claim 1, wherein the fertilizer has a nitrogen content of between 10-20 wt%.

10. The fertilizer of claim 1, wherein the fertilizer further comprises:

11. The fertilizer of claim 1, wherein the fertilizer further comprises:

12. The fertilizer of claim 1, wherein the fertilizer further comprises:

13. The fertilizer of claim 1, wherein the fertilizer further comprises:

14. The fertilizer of claim 1, wherein the fertilizer further comprises:

15. The fertilizer of claim 1, wherein the fertilizer further comprises:

and (5) growing a medium.

16. The fertilizer of claim 15, wherein the growth medium is selected from the group consisting of: soil, soilless growth media containing organic and/or inorganic components, organic soil amendments and mixtures thereof.

17. The fertilizer of claim 15, wherein the growth medium is selected from the group consisting of: soil, artificial plant growth substrates, organic soil conditioners, and mixtures thereof.

18. The fertilizer of claim 15, wherein the growth medium is selected from the group consisting of: soil, sand compost, peat, rice hulls, coconut fiber, coconut coir, polymer-based growth media, hydroponic nutrients and growth solutions, water, and mixtures thereof.

19. The fertilizer of claim 1, wherein the fertilizer further comprises:

a vehicle selected from the group consisting of water, aqueous solutions, slurries, and powders.

20. The fertilizer of claim 1, wherein the fertilizer further comprises:

an additive selected from the group consisting of: insecticides, fungicides, additional organic fertilizers, agricultural or horticultural adjuvants and mixtures thereof.

21. The fertilizer of claim 1, wherein the fertilizer further comprises:

an additive selected from the group consisting of: nematicides, additional organic fertilizers, biopesticides, adhesives, spreaders, surfactants, UV protectors and mixtures thereof.

22. The fertilizer of claim 1, wherein the fertilizer further comprises:

an additive selected from the group consisting of: nematicides, additional organic fertilizers, biopesticides, stickers, spreaders, dispersants, wetting agents, UV protectants and mixtures thereof.

23. The fertilizer of claim 1, wherein the fertilizer further comprises:

an additive selected from the group consisting of: bionematicides, additional organic fertilizers, adhesives, spreaders, surfactants, UV protectors and mixtures thereof.

24. The fertilizer of claim 1, wherein the fertilizer further comprises:

an additive selected from the group consisting of: bionematicides, additional organic fertilizers, stickers, spreaders, dispersants, wetting agents, UV protectants, and mixtures thereof.

25. The fertilizer of claim 1, wherein the fertilizer further comprises:

26. A fertilizer, comprising:

trichoderma viride NRRL B-50520 microorganism, and

a keratin substrate that is acted upon by the microorganism to release nitrogen.

27. The fertilizer of claim 26, further comprising:

28. The fertilizer of claim 26, further comprising:

a complementing microorganism in the form of a trichoderma species.

29. The fertilizer of claim 28, wherein the supplemental microorganism is selected from the group consisting of: trichoderma viride, Trichoderma harzianum, and Trichoderma atroviride, and combinations thereof.

30. The fertilizer of claim 26, further comprising:

31. The fertilizer of claim 26, wherein the fertilizer is in the form of granules, powder, slurry, film, and/or liquid suspension.

32. The fertilizer of claim 26, wherein the fertilizer is in the form of a liquid suspension comprising an amino acid and ammonia.

33. A method of enhancing plant growth, the method comprising:

providing a fertilizer comprising:

trichoderma viride NRRL B-50520 microorganism, and

a protein substrate that acts to release nitrogen by the microorganism; and is

34. The method of claim 33, wherein the protein substrate is selected from the group consisting of: feathers and feather meal.

35. The method of claim 33, further comprising:

a complementing microorganism in the form of a trichoderma species.

36. The method of claim 35, wherein the supplemental microorganism is selected from the group consisting of: trichoderma viride, Trichoderma harzianum, and Trichoderma atroviride, and combinations thereof.

37. The method of claim 33, further comprising:

38. The method of claim 33, wherein the fertilizer is in the form of granules, powder, slurry, film, and/or liquid suspension.

39. The method of claim 38, wherein the fertilizer is in the form of granules containing gelatin as a solidifying agent.

40. The method of claim 39, wherein the gelatin comprises 5-20% by weight of the granule.

41. The method of claim 38, wherein the fertilizer is in the form of a liquid suspension containing an amino acid and ammonia.

42. The method of claim 33, wherein the fertilizer has a nitrogen content of between 10-20 wt%.

43. The method of claim 33, wherein the fertilizer further comprises:

44. The method of claim 33, wherein the fertilizer further comprises:

a soluble nitrogen source selected from the group consisting of inorganic ammonia sources, fish meal or extracts, compost extracts, seaweed extracts, shrimp extracts, shellfish extracts, and combinations thereof.

45. The method of claim 33, wherein the fertilizer further comprises:

46. The method of claim 33, wherein the fertilizer further comprises:

47. The method of claim 33, wherein the fertilizer further comprises:

48. The method of claim 33, wherein the fertilizer further comprises:

49. The method of claim 33, wherein the fertilizer further comprises:

and (5) growing a medium.

50. The method of claim 49, wherein said contacting is performed before said plant or said plant seed is in said growth medium.

51. The method of claim 49, wherein said contacting is performed after said plant or said plant seed is in said growth medium.

52. The method of claim 49, wherein the growth medium is selected from the group consisting of: soil, soilless growth media containing organic and/or inorganic components, organic soil amendments and mixtures thereof.

53. The method of claim 49, wherein the growth medium is selected from the group consisting of: soil, artificial plant growth substrates, organic soil conditioners, and mixtures thereof.

54. The method of claim 49, wherein the growth medium is selected from the group consisting of: soil, sand compost, peat, rice hulls, coconut fiber, coconut coir, polymer-based growth media, hydroponic nutrients and growth solutions, water, and mixtures thereof.

55. The method of claim 33, wherein the fertilizer further comprises:

56. The method of claim 33, wherein the fertilizer further comprises:

57. The method of claim 33, wherein the fertilizer further comprises:

58. The method of claim 33, wherein the fertilizer further comprises:

59. The method of claim 33, wherein the fertilizer further comprises:

60. The method of claim 33, wherein the fertilizer further comprises:

61. The method of claim 33, wherein the fertilizer further comprises:

62. The method of claim 33, wherein the contacting is performed by: broadcast application, drop application, rotary application, liquid or dry in-furrow application, direct incorporation into soil or greenhouse planting mixtures, spray application, irrigation, injection, dusting, application of granules formed by extrusion or granulation, or coating the plant or the plant seed with the fertilizer.

63. The method of claim 33, wherein the plant growth enhancement is in the form of a larger root mass, a larger rooting depth, a larger shoot mass, a larger shoot leaf length, increased leaf greenness, increased yield, and improved clumping and vigor.

64. A method of manufacturing a fertilizer comprising an amino acid and ammonia, the method comprising:

providing a Trichoderma viride NRRL B-50520 microorganism;

providing a proteinaceous substrate which is acted upon by said microorganism to release nitrogen; and is

Contacting the microorganism with the proteinaceous substrate under conditions effective to cause the microorganism to exert an action on the proteinaceous substrate and produce a liquid suspension comprising amino acids and ammonia.

65. A fertilizer produced by the method of claim 64.

66. An isolated Trichoderma viride strain deposited at the culture collection of agricultural research services under accession number NRRL B-50520.

67. The isolated Trichoderma viride strain of claim 66 which is biologically pure.